Pyocyanin from Pseudomonas induces IL-8 production through the PKC and NF‑κB pathways in U937 cells

Chai,Wenshu; Zhang,Jia; Zhu,Zhitu; Liu,Wei; Pan,Dianzhu; Li,Ying; Chen,Baiyi

doi:10.3892/mmr.2013.1662

Pyocyanin from Pseudomonas induces IL-8 production through the PKC and NF‑κB pathways in U937 cells

Authors:
- Wenshu Chai
- Jia Zhang
- Zhitu Zhu
- Wei Liu
- Dianzhu Pan
- Ying Li
- Baiyi Chen
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Affiliations: Department of Respiratory Diseases, The First Affiliated Hospital, Liaoning Medical University, Jinzhou, Liaoning 121001, P.R. China, Hebei Medical University, Shijiazhuang, Hebei 050017, P.R. China, Department of Oncology, The First Affiliated Hospital, Liaoning Medical University, Jinzhou, Liaoning 121001, P.R. China, Department of Infectious Diseases, The First Affiliated Hospital of Chinese Medical University, Shenyang, Liaoning 110001, P.R. China
Published online on: August 30, 2013 https://doi.org/10.3892/mmr.2013.1662
Pages: 1404-1410

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Abstract

Pyocyanin, an extracellular product of Pseudomonas aeruginosa (P. aeruginosa), is important in invasive pulmonary infection. Pseudomonas infections are characterized by a marked influx of polymorphonuclear cells (neutrophils). An increased release of interleukin-8 (IL‑8), a potent neutrophil chemoattractant, in response to pyocyanin may contribute to the marked infiltration of neutrophils and subsequent neutrophil‑mediated tissue damage observed in Pseudomonas‑associated lung diseases. In order to explore the pathogenesis of pyocyanin on macrophages, we treated phorbol-12-myristate-13-acetate (PMA)-differentiated U937 cells with pyocyanin in vitro and explored the expression of IL‑8 using reverse transcription polymerase chain reaction (RT‑PCR) and enzyme‑linked immunosorbent assay (ELISA). In the present study, we also investigated the activation of protein kinase C (PKC) and nuclear factor (NF)‑κB in PMA-differentiated U937 cells by western blotting and immunohistochemical methods. It was found that pyocyanin increased IL‑8 release and mRNA expression in differentiated U937 cells in a concentration- and time-dependent manner. Calphostin C (Cal C), a protein kinase C (PKC) inhibitor, and pyrrolidine dithiocarbamate (PDTC) , an NF‑κB inhibitor, blocked IL‑8 expression in a concentration-dependent manner in pyocyanin‑induced U937 cells. We concluded that pyocyanin promotes IL‑8 secretion and mRNA expression in a concentration- and time-dependent manner and furthermore, that the PKC and NF‑κΒ signaling pathways may be involved in the expression of IL‑8 in pyocyanin‑infected PMA-differentiated U937 cells.

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